Immunocytochemistry/Immunofluorescence analysis of Cytohesin 2 in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2728 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Cytohesin 2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunocytochemistry/Immunofluorescence analysis of Cytohesin 2 in HepG2 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2728 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Cytohesin 2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunocytochemistry/Immunofluorescence analysis of Cytohesin 2 in NIH-3T3 cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2728 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Cytohesin 2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) was performed on normal biopsies of deparaffinized human colon carcinoma tissue. Heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and incubated with ab2728 (1:50) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) was performed on normal biopsies of deparaffinized human stomach tissue. Heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and incubated with ab2728 (1:50) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.